LOCUS       pLMPd-Ametrine.        6740 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Retroviral vector with an mAmetrine marker, for expressing a short 
            hairpin RNA (shRNA) with an """"UltramiR"""" microRNA scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    pLMPd-Ametrine
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6740)
  AUTHORS   transOMIC
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6740)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..515
                     /label=MSCV
                     /note="Murine embryonic stem cell virus promoter including
                     the enhancer and promoter region of PCMV virus and 5' 
                     untranslated region of dl-587rev retrovirus"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
     ncRNA           1443..1564
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     misc_RNA        1565..1633
                     /label=shRNA
                     /note="shRNA"
                     /note="short hairpin RNA"
     gap             1568..1589
                     /estimated_length=22
     gap             1609..1630
                     /estimated_length=22
     ncRNA           1634..1760
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     promoter        1774..2273
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             2294..3010
                     /label=mAmetrine1.1
                     /note="photostable yellow fluorescent protein with a large 
                     Stokes shift, excitable with violet light (Ai et al., 
                     2008)"
     LTR             3102..3616
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem
                     cell virus"
     primer_bind     complement(3785..3801)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3809..3825)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3833..3863)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3878..3899)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4187..4775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4949..5806)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5807..5911)
                     /label=AmpR promoter
     LTR             6739..6740
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctctcga ctagggataa cagggtaatt
     1441 gtttgaatga ggcttcagta ctttacagaa tcgttgcctg cacatcttgg aaacacttgc
     1501 tgggattact tcttcaggtt aacccaacag aaggctaaag aaggtatatt gctgttgaca
     1561 gtgagcgnnn nnnnnnnnnn nnnnnnnnnt agtgaagcca cagatgtann nnnnnnnnnn
     1621 nnnnnnnnnn tgcctactgc ctcggacttc aaggggctac tttaggagca attatcttgt
     1681 ttactaaaac tgaatacctt gctatctctt tgatacattt ttacaaagct gaattaaaat
     1741 ggtataaatt aaatcacttt tttcaattct accgggtagg ggaggcgctt ttcccaaggc
     1801 agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct
     1861 ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc
     1921 ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg
     1981 cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca
     2041 gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct
     2101 ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg
     2161 ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc
     2221 aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca
     2281 gcccaagctt accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2341 ggtcgagctg gacggcgacg taaacggcca caggttcagc gtgcgcggcg agggcgaggg
     2401 cgatgccacc aacggcaagc tgaccctgaa gttcatctgc acctccggcg agctgcccgt
     2461 gccctggccc accctcgtga ccaccctgtc ttacggcgtg cagtgcttcg cccgctaccc
     2521 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2581 gcgcaccatc tccttcaagg acgacggcag ctacaggacc cgcgccgagg tgaagttcga
     2641 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2701 catcctgggg cacaagctgg agtacaacat gaacgcgtgg gacgcgtata tcacggccga
     2761 caagcagaag aacggcatca aagcgaactt caagatcgag cacaacgtca aggacggcgg
     2821 cgtgcagctc gccgacgcgt accagcagaa cacccccatc ggcgacggct ccgtgctgct
     2881 gcctgacaac cactacctga gcttccagag caagctgttc aaagacccca acgagcagcg
     2941 cgatcacatg gtcctgctgg agttcgttac cgccgccggg ataactcccg gcatggacga
     3001 gctgtacaag taaagcggcc gcgactctag agtcgacctg caggcatgca agcttatcga
     3061 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     3121 ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag
     3181 aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg
     3241 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg
     3301 cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac
     3361 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
     3421 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt
     3481 cctccgatag actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca
     3541 tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc
     3601 agcgggggtc tttcatgggt aacagtttct tgaagttgga gaacaacatt ctgagggtag
     3661 gagtcgaata ttaagtaatc ctgactcaat tagccactgt tttgaatcca catactccaa
     3721 tactcctgaa atagttcatt atggacagcg cagaagagct ggggagaatt aattcgtaat
     3781 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     3841 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     3901 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3961 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4021 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4081 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4141 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4201 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4261 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4321 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4381 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4441 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4501 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4561 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4621 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4681 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4741 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4801 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4861 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4921 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4981 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5041 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5101 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5161 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5221 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5281 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5341 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5401 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5461 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5521 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5581 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5641 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5701 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5761 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5821 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5881 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5941 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     6001 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     6061 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     6121 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     6181 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     6241 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     6301 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     6361 cagggttttc ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg catgcaagga
     6421 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     6481 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     6541 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggcg
     6601 attagtccaa tttgttaaag acaggatatc agtggtccag gctctagttt tgactcaaca
     6661 atatcaccag ctgaagccta tagagtacga gccatagata aaataaaaga ttttatttag
     6721 tctccagaaa aaggggggaa
//
